TNFα promotes motility and lamellipodia dynamics of infected macrophages.

<p><b>A</b>) Left: Quantification of TNFα concentration in culture supernatants of TaC12, Thei and TaH12810 cells by ELISA. Red line: detection limit (0.078 ng/ml). Right: Quantification of TNFα mRNA expression in TaC12, Thei and TaH12810 cells by qRT-PCR. <b>B</b>). Quantification of TNFα concentration in culture supernatants of parasitized or BW720c-cured TaC12, Thei and TaH12810 cells by ELISA. <b>C</b>) Quantification of TNFα mRNA expression in parasitized or BW720c-cured TaC12, Thei and TaH12810 cells by qRT-PCR. <b>D</b>) Quantification of path lengths of individual TaC12 and Thei cells migrating for 6 and 12 h, respectively, −/+ TNFα (25 ng/ml, TaC12 n = 31, 40, Thei n = 90, 90). <b>E</b>) Matrigel invasion assay: Quantification of TNFα-induced transmigration of TaC12 cells in 24 h relative to unstimulated controls. <b>F</b>) Left IF images: Visualization of TNFα (25 ng/ml, 24 h) effects on F-actin-rich protrusions in 2D (upper) and inside matrigel (3D, lower). Magnifications are 4× of boxed areas. Right, box plots: Means of relative areas (ratio lamellipodia area∶cell area) of lamellipodia in TaC12 and Thei cells that were quantified as depicted in F upper (TaC12: n = 77, 38, Thei: n = 30, 30). <b>G</b>) Mean percentage of cells with lamellipodia in TaC12 and Thei cells treated with BW720c and TNFα as indicated. See also <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1004003#ppat.1004003.s001" target="_blank">Figure S1</a>.